MicroRNAs (miRNAs) are regulators of gene expression that often suppress the translation of protein coding mRNAs through a RNA-induced silencing complex (RISC). Although some methods of identifying miRNA targets are available (e.g. luciferase assays) these methods are laborious, expensive and imperfect.
Methods of identifying miRNA targets is described in articles entitled “MicroRNA-10a Binds the 5′UTR of Ribosomal Protein mRNAs and Enhances Their Translation” by Orom et al. (Mol. Cell 30, 460-71, 2008) and “Isolation of microRNA targets using biotinylated synthetic microRNAs” by Orom et al. (Methods 43, 162-5, 2007). Also see an article entitled “Profiling Direct mRNA-microRNA interactions using synthetic biotinylated microRNA-duplexes” by Wani et al. (printed online at BioRxIV, May 22, 2014). These techniques are referred to as RNA pull-down assays. Briefly, a miRNA of interest in synthesized with a 3′biotin group. The miRNA binds to its mRNA targets in cells. The resulting complex is separated from other cellular materials by, for example, streptavidin beads, purified and subjected to PCR, microarray or sequencing analysis to identify the molecular targets of the miRNA.
The discussion above is merely provided for general background information and is not intended to be used as an aid in determining the scope of the claimed subject matter.